China Animal Husbandry and Veterinary Medicine ›› 2025, Vol. 52 ›› Issue (2): 874-889.doi: 10.16431/j.cnki.1671-7236.2025.02.037

• Basic Veterinary Medicine • Previous Articles    

Exploration of the Mechanism of Action of Angelica Decoction on Atopic Dermatitis Mice Using UPLC-Q-TOF-MS Combined with Network Pharmacology

HUANG Yongxi1, YAN Pupu1, GUO Weili2, LI Yana3, ZHU Jun1, LIU Man1, HOU Chaoqun1, QIN Meilin1, LI Rong1, WU Yi4, SU Yingbing1, YANG Xiaolin1, GUO Liwei1, WANG Xiong5, DAI Gang5   

  1. 1. Engineering Research Center of Ecology and Agricultural Use of Wetland, Ministry of Education, College of Animal Science and Technology, Yangtze University, Jingzhou 434000, China;
    2. No. 6 Mildle School of Shahe, Shahe 054100, China;
    3. Yichang Renfu Pharmaceutical Co., Ltd., Yichang 443000, China;
    4. College of Veterinary Medicine, Yunnan Agricultural University, Kunming 650500, China;
    5. Jingzhou Mingde Technology Co., Ltd., Jingzhou 434001, China
  • Received:2024-09-02 Published:2025-01-24

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Abstract: 【Objective】 This study was aimed to investigate the effects of Angelica decoction (DGYZ) on the skin lesions of mice with atopic dermatitis (AD) induced by 2,4-dichloronitrobenzene (DNCB),provide a reference for the clinical diagnosis and treatment of AD in pets. 【Method】 A total of 90 BALB/c mice were selected and divided into six groups:Control group,DNCB group,low-dose (9.8 mg/kg DGYZ),medium-dose (19.6 mg/kg DGYZ),high-dose (39.3 mg/kg DGYZ) DGYZ groups and cetirizine group (4 mg/kg cetirizine),with 15 mice in each group.The mice in control group were treated with acetone solution as a comparison,while mice in the other groups were subjected to repeated application of DNCB on their backs to establish a mouse model of AD,followed by drug treatment according to their respective groups.After the treatment,skin samples from the back,blood and spleens were collected from the mice for subsequent analysis.Tissue thickness and the number of mast cells were observed through HE and toluidine blue staining.ELISA was used to measure serum immunoglobulin E (IgE) level.The contents of white blood cells (WBC),neutrophils and eosinophils in the blood were measured by automatic blood cell analyzer.Flow cytometry was used to detect CD4+/CD8+ cell differentiation and Th1/2 cell differentiation.Real-time quantitative PCR was used to measure the expression of cytokine-related mRNA in the back skin of mice.Ultra-high-performance liquid chromatography-time-of-flight mass spectrometer (UPLC-TOF-MS) was used to analyze the main active ingredients of DGYZ,combined with network pharmacology to explore the main targets and mechanisms of action of DGYZ in the treatment of AD.Finally,the mRNA expression of JAK1-STAT3 signaling pathway was detected by Real-time quantitative PCR. 【Result】 Compared with control group, mice in DNCB group exhibited severe skin lesions and epidermal thickening on their backs, along with a significant increase in the number of leukocytes, neutrophils, and eosinophils in the blood (P＜0.05). Compared with DNCB group,the epidermal thickening of mice in DGYZ groups at various doses and cetirizine group was effectively alleviated.And the counts of white blood cells,neutrophils and eosinophils were all decreased compared with DNCB group,DGYZ high-dose group and cetirizine groups were significantly different (P＜0.05).In addition,the serum IgE expression of mice in DGYZ high-dose group and cetirizine group were significantly reduced (P＜0.05).Flow cytometry showed that the balance of CD4+/CD8+ cells and Th1/2 cells was disrupted in DNCB group,and different doses of DGYZ significantly improved this situation (P＜0.05).Real-time quantitative PCR detection results showed that compared with blank group,the mRNA expression of interferon gamma (IFN-γ) and interleukin-12 (IL-12) genes in the skin tissue of mice in DNCB group were significantly downregulated (P＜0.05).The mRNA expression of IL-4,IL-13 and IL-6 genes were significantly increased (P＜0.05).Compared with DNCB group,the mRNA expression of IFN-γ and IL-12 genes in skin tissue were gradually increased after feeding DGYZ treatment,while the mRNA expression of IL-4,IL-13,and IL-6 genes were significantly decreased (P＜0.05).UPLC-TOF-MS identified 89 ingredients of DGYZ,including 64 active ingredients.The results of network pharmacology analysis showed that the above 64 active ingredients exerted therapeutic effects on AD-like skin lesions by acting on genes such as signal transduction and transcriptional activation protein 3 (STAT3),protein kinase B (AKT1),as well as JAK/STAT,NF-κB and Th17 cell differentiation signaling pathways.Real-time quantitative PCR detection results showed that DNCB could induce the activation of JAK1-STAT3 signaling pathway in mice,while DGYZ could inhibit the activation of this pathway. 【Conclusion】 DGYZ could alleviate AD-like skin lesions by reducing the number of mast cells in skin and the expression of IgE in serum of mice,regulating the immune balance,reducing the mRNA expression of pro-inflammatory factors and inhibiting the activation of JAK1-STAT3 signaling pathway.The results could provide a theoretical and experimental basis for the treatment of AD-like skin lesions in animals with traditional Chinese medicine.

Key words: Angelica decoction (DGYZ); AD-like skin lesions; 2,4-dichloronitrobenzene (DNCB); flow cytometry; network pharmacology

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